Transcription factors play crucial regulatory role within the cell and the entire multicellular organism. The important factor is its ability to interact with other regulatory proteins and DNA. Despite the fact that a large part of the interaction network is already documented, detailed information on the structure and dynamics of protein-protein and protein-DNA complexes is still scarce. In this thesis we focused on the possibility of studying conformational changes given by the transcription factor-DNA complex formation using the methods of structural mass spectrometry: hydrogen/deuterium exchange and chemical crosslinking. As a model, we chose a transcription factor FOXO4 which DNA binding domain is structurally well characterized both in free form and in the complex with DNA.

Proteins of the CSL family (CBF1/RBP-Jκ/Suppressor of Hairless/LAG-1) act as effectors of the Notch signalling pathway in metazoan organisms. They function as repressors or activators of gene transcription in the framework of this pathway and influence many developmental processes. Metazoan CSL proteins can regulate gene expression Notch-independently as well. Notch-independent functions of CSL proteins might be evolutionarily ancestral and in cells and organisms may be important equally as Notch-dependent functions. Presence of CSL proteins was identified in several fungal species, organisms lacking the Notch signalling pathway components and most of known metazoan interacting partners of CSL proteins. CSL paralogs of the fission yeast Schizosaccharomyces pombe, cbf11 and cbf12, are non-essential genes encoding proteins localized in the nucleus of the cell. They exert antagonistic effects on regulation of processes like coordination of nuclear and cellular division and cell cycle progression, ploidy maintenance, cell adhesion and other. In this study, we have proved that both CSL paralogs are able to sequence-specifically bind the CSL-response element DNA in vitro and Cbf11 in vivo as well. Both proteins could activate gene expression in vivo and perform the function of transcription factors....

Loss of hearing affects more than 10 % of the population, and one newborn in a thousand is born with defects of the inner ear. Transcriptional factors involved in the development of inner ear are important in our understanding of the causes of inner ear defects. ISLET1 is one of these factors. ISLET1 expression is detected in the sensory and neuronal cells of the inner ear. It participates in otocyst formation, and the specification and differentiation of cells of cochlea and vestibular system. The functional role of ISLET1 during inner ear development was investigated. Its role was studied by using Pax2-Isl1 transgenic mice that overexpress Islet1 under the control of the Pax2 promoter. Two transgenic lines were generated, Pax2-Isl1/300 and Pax2- Isl1/52. Two copies of the Pax2-Isl1 transgene were inserted to Pax2-Isl1/300 genome and one copy was inserted to the Pax2-Isl1/52 genome. Defects in sense of hearing were detected in both lines and circling behavior, a defect of balance, was detected in the Pax2-Isl1/300 transgenic mice. We observed high postnatal lethality in heterozygote transgenic mice. Pax2-Isl1/52 homozygote mutation is lethal at embryonic day 10 (E10,5). Pax2-Isl1/300 homozygote letality couldn't be detected because of the inability to breed heterozygote mutated mice of this line....

Title: Study of proton-proton and heavy-ion collisions with the ATLAS experi- ment at the LHC Author: Mgr. Petr Balek Department: Institute of Particle and Nuclear Physics Supervisor: Doc. RNDr. Jiří Dolejší, CSc. Abstract: The presented results show a suppression in a production of charged hadrons in heavy-ion collisions with respect to the production measured in proton- proton collisions. Lead-lead data at √ sNN = 2.76 TeV were recorded by the AT- LAS experiment at the LHC in 2010 and 2011 and they have a total integrated luminosity of 0.15 nb−1 , and proton-proton data at √ s = 2.76 TeV were recorded in 2011 and 2013 having an integrated luminosity of 4.2 nb−1 . The results are presented in wide transverse momentum (0.5 < pT < 150 GeV) and pseudorapid- ity (|η| < 2) ranges, and also in different centrality intervals of Pb+Pb collisions. Nuclear modification factor RAA shows a clear pT-dependence with a minimum at around 7 GeV, however above 60 GeV, it reaches a plateau with no significant pT-dependence. As a function of |η|, the RAA is flat over whole pT region. Keywords: ATLAS detector, heavy-ion collisions, charged-hadron spectra, nuclear modification factor RAA

In Introduction, the author of this dissertation deals with postoperative venous thromboembolism (VTE), hereditary and acquired risk factors, prophylaxis regimens and recent recommendation of VTE prevention in surgery. In Practical part of this work the author assesses the risk of VTE in surgical patients according to risk assessment model. Genetic testing is carried out in all patients to determine the incidence of hereditary thrombophilia and coagulation markers are measured in 28-days postoperative period. Prevalence of VTE in 1-year postoperative period is observed. The results are analysed in group of patients with benign disease (hernia and gallstone disease) and group of patients with malignancy (colorectal cancer and pancreatic cancer) separately. The objective of this work is to determine the incidence of the most frequent thrombophilic mutations (factor V Leiden mutation and protrombin G20210A mutation) and assess the impact of hereditary thrombophilia on incidence of postoperative venous thromboembolism in general surgery. Validation of venous thrombosis risk assessment model recommended by Czech Society for Thrombosis and Hemostasis is further objective.

This paper deals with the expression of vascular endothelial growth factor (vascular endothelial growth factor, VEGF) and its use in tissue engineering of vascular wall. During the work interaction of endothelial cells with the modified fibrin-based biomaterial into which vascular endothelial growth factor (VEGF-A121) has been incorporated was monitored. This modification supported the adhesion and growth of endothelial cells. Vascular endothelial growth factor VEGF-A121 is signal glycoprotein that activates transmembrane receptors on endothelial cells. VEGF-A121 is a key regulator in vasculogenesis, angiogenesis, proliferation, migration and survival of endothelial cells. In this work, this protein was heterologously expressed at a thioredoxin fusion partner in an expression system of E. coli Origami B (DE3). Recombinant VEGF-A121 was additionally coexpressed with bacterial chaperones GroEL/GroES for potential increase of its solubility and biological activity. In the next part of this work thin fibrin network was prepared by catalytic action of thrombin on the polystyrene-bound monolayer of fibrinogen. This network has been further enriched by vascular endothelial growth factor (VEGF-A121), which was covalently incorporated in it by enzyme activity of transglutaminase (factor XIIIa). The last...

Title: Some aspects of low-energy QCD at the precision frontier Author: Tomáš Husek Department/Institute: Institute of Particle and Nuclear Physics Supervisor of the doctoral thesis: doc. RNDr. Karol Kampf, Ph.D. Abstract: This thesis concentrates on some low-energy aspects of QCD, namely on those which are connected to the electromagnetic decays of lightest neutral pseudoscalar mesons. Calculations of radiative corrections to neutral pion decays (the Dalitz decay and the rare decay) and a novel model for the pion electro- magnetic transition form factor are subjects discussed in the attached papers, which this work is based on. The associated theoretical aspects including Chiral Perturbation Theory or the large-Nc limit are introduced. We also discuss the complications which arise when the calculations of radiative corrections for η(′) Dalitz decays are performed. Some details about the collaboration with experi- ments which incorporate the calculation of the published corrections are provided. Last but not least, some techniques related to loop integrals are shown. Keywords: Chiral Perturbation Theory, large-Nc limit, radiative corrections, pion electromagnetic transition form factor 1

Ziel der vorliegenden Arbeit war die Entwicklung eines Sensors zur gleichzeitigen Bestimmung von Spannungsintensitätsfaktoren (K-Faktoren) und der Risslage. Im Gegensatz zum Stand der Technik werden zwei Aufgaben zur Charakterisierung der Risse (Detektierung von Risslage und Größe und Quantifizierung des Beanspruchungszustandes an der Rissspitze) mit Hilfe nur eines Sensors ermöglicht. Es wurde effiziente, kostengünstige und leicht zu applizierende Technik zur experimentellen bruchmechanischen Beanspruchungsanalyse entwickelt. Eine Struktur mit Riss wird großflächig mit einer piezoelektrischen Polyvinylidenfluorid-Folie (PVDF-Folie) beklebt und eine ausreichende Anzahl von Elektroden befindet sich im Bereich des möglichen/vorhandenen Risses. Die an den Elektroden abgegriffenen elektrischen Potentiale bzw. Ladungen sind ein Maß für die Verzerrungen an der Oberfläche des Bauteils und ermöglichen die Berechnung der bruchmechanischen Größen am Riss. Dabei beschränkte man sich auf die Anwendung des Sensorkonzeptes im Rahmen der Scheiben- und Plattentheorie.

Dita Kašparová Chronická hypoxie a exprese genů 4 Abstract Adaptation to chronic hypoxia (CH) is characterized by a variety of functional changes in order to maintain metabolic and energy homeostasis. It has been known for many years that both humans and animals indigenous or adapted to high-altitude hypoxia are more tolerant to an acute ischemic injury of the heart. Cardioprotective mechanisms activated by adaptive responses to chronic hypoxia can be the result of altered transcriptional regulations in left ventricles. Here we report results from the gene expression profiling of adaptive responses in three models of chronically hypoxic heart. Adult male Wistar rats were exposed for 21 days to either continuous normobaric hypoxia (CCH; 10% O2) or CCH interrupted daily by 1-hour reoxygenation (RCH) or CCH interrupted daily by 16-hour (CIH). Cardiprotective effect of CCH adaptation is abolished by brief daily reoxygenation, RCH adaptation. In the present study, we aimed to determine myocardial mRNA expression of 19 candidate genes divided into three important groups: i) Hypoxia inducible factor (HIF1α) and its prolyl and asparaginyl hyroxylases (PHDs and FIH respectively, ii) Creatine kinase (CK) isoenzymes which play important role in energy homeostases of heart and iii) the group of main enzymatic...

Optimization of biologic therapy in children with inflammatory bowel disease (IBD) using modern biomarkers Abstract to thesis Study programme: Biochemistry and Pathobiochemistry Introduction: In adults, infliximab (IFX) levels correlate with disease activity and antibodies to IFX (ATIs) predict treatment failure. We aimed to determine the association of IFX levels and ATIs with disease activity in paediatric population. Methods: This study was performed as a prospective observational study. We prospectively collected blood, stool, and clinical data from 65 patients (age 10.5-15.1 years) with Crohn's disease (CD) before IFX administration, and measured IFX trough levels, ATIs, and faecal calprotectin levels (CPT). We used multivariate analysis to identify the predictors of IFX levels. IFX and ATIs levels were meassured using ELISA. Results: Lower levels of IFX were associated with ATIs positivity (OR [odds ratio] 0.027, CI [confidence interval] 0.009-0.077). Higher C-reactive protein (CRP) level, erythrocyte sedimentation rate (ESR) and CPT levels were found in patients with lower IFX levels. The optimal combination of specificity (50%) and sensitivity (74%) for disease activity was calculated for IFX levels ≥ 1.1 µg/ml using CRP level < 5 mg/l as a marker of laboratory remission. In a model that used CPT ≤...